A Method to Study the C924T Polymorphism of the Thromboxane A2 Receptor Gene

J Vis Exp. 2019 Apr 1:(146). doi: 10.3791/57289.

Abstract

The thromboxane A2 receptor (TBXA2R) gene is a member of the G-protein coupled superfamily with seven-transmembrane regions. It is involved in atherogenesis progression, ischemia, and myocardial infarction. Here we present a methodology of patient genotyping to investigate the post-transcriptional role of the C924T polymorphism (rs4523) situated at the 3' region of the TBXA2 receptor gene. This method relies on DNA extraction from whole blood, polymerase chain reaction (PCR) amplification of the TBXA2 gene portion containing the C924T mutation, and identification of wild type and/or mutant genotypes using a restriction digest analysis, specifically a restriction fragment length polymorphism (RFLP) on agarose gel. In addition, the results were confirmed by sequencing the TBXA2R gene. This method features several potential advantages, such as high efficiency and the rapid identification of the C924T polymorphism by PCR and restriction enzyme analysis. This approach allows a predictive study for plaque formation and atherosclerosis progression by analyzing patient genotypes for the TBXA2R C924T polymorphism. Application of this method has the potential to identify subjects who are more susceptible to atherothrombotic processes, in particular subjects in a high-risk, aspirin-treated group.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Base Sequence
  • Genotype
  • Humans
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length / genetics
  • Polymorphism, Single Nucleotide / genetics*
  • Receptors, Thromboxane A2, Prostaglandin H2 / genetics*
  • Restriction Mapping

Substances

  • Receptors, Thromboxane A2, Prostaglandin H2